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Methane is the second most important greenhouse gas contributing to about 20% of global warming. Its mitigation is conducted by methane oxidizing bacteria that act as a biofilter using methane as their energy and carbon source. Since their first discovery in 1906, methanotrophs have been studied using a complementary array of methods. One of the most used molecular methods involves PCR amplification of the functional gene marker for the diagnostic of copper and iron containing particulate methane monooxygenase. To investigate the diversity of methanotrophs and to extend their possible molecular detection, we designed a new set of degenerate methane monooxygenase primers to target an 850 nucleotide long sequence stretch from  pmoC  to  pmoA.  The primers were based on all available full genomic  pmoCAB  operons. The newly designed primers were tested on various pure cultures, enrichment cultures and environmental samples using PCR. The results demonstrated that this primer set has the ability to correctly amplify the about 850 nucleotide long  pmoCA  product from  Alphaproteobacteria ,  Gammaproteobacteria ,  Verrucomicrobia  and the NC10 phyla methanotrophs. The new primer set will thus be a valuable tool to screen ecosystems and can be applied in conjunction with previously used  pmoA  primers to extend the diversity of currently known methane-oxidizing bacteria.   Electronic supplementary material  The online version of this article (doi:10.1186/s13568-017-0466-2) contains supplementary material, which is available to authorized users.

Survey of methanotrophic diversity in various ecosystems by degenerate methane monooxygenase gene primers